Flexible and rapid construction of viral chimeras applied to Hepatitis C Virus

A novel and broadly applicable strategy combining site directed mutagenesis and DNA assembly for constructing seamless viral chimeras is described using Hepatitis C Virus as an exemplar. Full-length HCV genomic cloning cassettes, which contained flexibly situated restriction endonuclease sites, were prepared via a single site-directed mutagenesis reaction and digested to receive PCR amplified virus envelope genes by In-Fusion cloning. Using this method we were able to construct gene-shuttle cassettes for generation of cell culture-infectious JFH-1-based chimeras containing genotype 1-3 E1E2 genes. Importantly, using this method we also show that E1E2 clones that were not able to support cell entry in the HCV pseudoparticle assay did confer entry when shuttled into the chimeric cell culture chimera system. This method can be easily applied to other genes of study and other viruses and, as such, will greatly simplify reverse genetics studies of variable viruses

Cd98hc (slc3A2) sustains amino acid and nucleotide availability for cell cycle progression

CD98 heavy chain (CD98hc) forms heteromeric amino acid (AA) transporters by interacting with different light chains. Cancer cells overexpress CD98hc-transporters in order to meet their increased nutritional and antioxidant demands, since they provide branched-chain AA (BCAA) and aromatic AA (AAA) availability while protecting cells from oxidative stress. Here we show that BCAA and AAA shortage phenocopies the inhibition of mTORC1 signalling, protein synthesis and cell proliferation caused by CD98hc ablation. Furthermore, our data indicate that CD98hc sustains glucose uptake and glycolysis, and, as a consequence, the pentose phosphate pathway (PPP). Thus, loss of CD98hc triggers a dramatic reduction in the nucleotide pool, which leads to replicative stress in these cells, as evidenced by the enhanced DNA Damage Response (DDR), S-phase delay and diminished rate of mitosis, all recovered by nucleoside supplementation. In addition, proper BCAA and AAA availability sustains the expression of the enzyme ribonucleotide reductase. In this regard, BCAA and AAA shortage results in decreased content of deoxynucleotides that triggers replicative stress, also recovered by nucleoside supplementation. On the basis of our findings, we conclude that CD98hc plays a central role in AA and glucose cellular nutrition, redox homeostasis and nucleotide availability, all key for cell proliferation

Tratamientos psicológicos empíricamente apoyados para adultos: Una revisión selectiva

Antecedentes: los tratamientos psicológicos han mostrado su eficacia, efectividad y eficiencia para el abordaje de los trastornos mentales; no obstante, considerando el conocimiento científico generado en los últimos años, no se dispone de trabajos de actualización en español sobre cuáles son los tratamientos psicológicos con respaldo empírico. El objetivo fue realizar una revisión selectiva de los principales tratamientos psicológicos empíricamente apoyados para el abordaje de trastornos mentales en personas adultas. Método: se recogen niveles de evidencia y grados de recomendación en función de los criterios propuestos por el Sistema Nacional de Salud de España (en las Guías de Práctica Clínica) para diferentes trastornos psicológicos. Resultados: los resultados sugieren que los tratamientos psicológicos disponen de apoyo empírico para el abordaje de un amplio elenco de trastornos psicológicos. El grado de apoyo empírico oscila de bajo a alto en función del trastorno psicológico analizado. La revisión sugiere que ciertos campos de intervención necesitan una mayor investigación. Conclusiones: a partir de esta revisión selectiva, los profesionales de la psicología podrán disponer de información rigurosa y actualizada que les permita tomar decisiones informadas a la hora de implementar aquellos procedimientos psicoterapéuticos empíricamente fundamentados en función de las características de las personas que demandan ayuda. Background: Psychological treatments have shown their efficacy, effectiveness, and efficiency in dealing with mental disorders. However, considering the scientific knowledge generated in recent years, in the Spanish context, there are no updating studies about empirically supported psychological treatments. The main goal was to carry out a selective review of the main empirically supported psychological treatments for mental disorders in adults. Method: Levels of evidence and degrees of recommendation were collected based on the criteria proposed by the Spanish National Health System (Clinical Practice Guidelines) for different psychological disorders. Results: The results indicate that psychological treatments have empirical support for the approach to a wide range of psychological disorders. These levels of empirical evidence gathered range from low to high depending on the psychological disorder analysed. The review indicates the existence of certain fields of intervention that need further investigation. Conclusions: Based on this selective review, psychology professionals will be able to have rigorous, up-to-date information that allows them to make informed decisions when implementing empirically based psychotherapeutic procedures based on the characteristics of the people who require help

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p<0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p<0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised

Soy Niña

Este libro pretende contribuir al reencuentro de la educación con esas finalidades que verdaderamente importan a una niña o un niño: ser feliz, jugar, vivir juntos y (no) aprender. Para ello hemos puesto el arte, nuestras experiencias y el saber acumulado al servicio del disfrute, el cuestionamiento, el análisis crítico y la construcción común de un presente deseable. Un texto colaborativo coordinado por Ignacio Calderón Almendros y realizado por alumnado de Educación y Cambio Social en el Grado en Educación Infantil de la Universidad de Málaga

CD98hc orchestrates amino acid and glucose availability, redox homeostasis and energetic and nucleotide metabolism = CD98hc sostiene la disponibilidad de aminoácidos y glucosa, la homeóstasis óxido-reducción y el metabolismo energético y de nucleótidos

[eng] The wide variety of disorders associated with the malfunction of amino acid (AA) transporters reflect the relevant roles they fulfil in human physiology. Heteromeric AA transporters (HATs), one of the eleven families of AA plasma membrane transporters in human cells, are formed by an ancillary heavy subunit, which brings the holotransporter to the plasma membrane and a catalytic light subunit. The heavy subunit CD98hc (also named 4F2hc or SLC3A2) can dimerize with any of six light subunits (LAT1, LAT2, xCT, y+LAT1, y+LAT2 and asc1) that confer substrate specificity to the heterodimer. Cancer cells overexpress CD98hc-transporters in order to meet their increased nutritional and antioxidant demands, since these transporters are crucial to control reactive oxygen species and intracellular AA levels, thus sustaining cell survival and proliferation. CD98hc-xCT is required for cystine uptake, which is rapidly reduced to cysteine inside the cell, a rate-limiting AA in the synthesis of reduced glutathione (GSH). Since GSH is vital for controlling the levels of reactive oxygen species in the cell, loss of CD98hc-xCT in vitro cannot be compensated, leading to cell death by ferroptosis. Supplementation of culture media with β-mercaptoethanol rescues CD98hc-deficient cell survival. Under such conditions cells lacking CD98hc still present increased levels of reactive oxygen species and intracellular AA imbalance, probably due to the loss of CD98hc-LAT1. Consequently, cells lacking CD98hc present alterations in the stress-sensing signalling pathways mTORC1 and Integrated Stress Response- mediated by eIF2α, leading to defective protein synthesis and proliferation. Furthermore, CD98hc regulates the energy metabolism of the cell. Indeed, lack of CD98hc triggers a reduction the glucose uptake and glucose-related metabolic pathways, such as glycolysis and the pentose phosphate pathway, forcing cells to use mitochondrial oxidative metabolism in order to maintain the cellular ATP levels, suggesting a metabolic rewiring. The abrogation of the pentose phosphate pathway flux by CD98hc ablation triggers a dramatic reduction in the levels of ribose-5P, which forms the sugar backbone of all nucleotides. Consequently, these cells present a dramatic reduction in the nucleotide pool, which cause replicative stress, enhanced DNA Damage Response, delayed cell cycle progression during the S-phase and diminished rate of mitosis, all recovered by nucleoside supplementation. In addition, reduction in the intracellular content of the essential branched-chain AAs (leucine, isoleucine and valine) and aromatic AAs (tyrosine, phenylalanine and tryptophan), all transported by CD98hc-LAT1, leads to repressed expression of RRM2, the only enzyme able to catalyse the reduction of ribonucleotides to deoxyribonucleotides, resulting as well in replicative stress. Taking all of these considerations, CD98hc may be a putative target in pathophysiological scenarios, thus, generating drugs against CD98hc could represent a novel option, widening the therapeutic window of cancer therapy by inducing an immediate impairment in: the redox counterbalance capacity, the intracellular AA content for proper protein synthesis and cell proliferation and the pentose phosphate pathway flux required for producing ribose-5P to sustain nucleotide biosynthesis of the cells.[spa] La gran variedad de trastornos asociados con el mal funcionamiento de los transportadores de aminoácidos (AA) evidencia la importancia de las funciones que éstos llevan a cabo en la fisiología humana. Los transportadores heteroméricos de AAs (HATs) están formados por una subunidad pesada, que sitúa el heterodímero en la membrana plasmática, y una subunidad ligera que confiere la capacidad catalítica al transportador. La subunidad pesada CD98hc (también denominada 4F2hc o SLC3A2) puede dimerizar las subunidades ligeras LAT1, LAT2, xCT, y + LAT1, y + LAT2 y asc1, confiriendo especificidad de sustrato al heterodímero. Las células tumorales sobreexpresan los transportadores asociados a CD98hc para satisfacer sus demandas nutricionales y antioxidantes, ya que estos transportadores son cruciales para controlar las especies reactivas de oxígeno y los niveles intracelulares de AA, soportando la viabilidad y la proliferación celular. CD98hc-xCT es el responsable de la captación de cistina, necesaria para sintetizar glutatión y por tanto para mantener la homeostasis óxido-reducción. En la célula. Por este motivo, la eliminación de CD98hc-xCT in vitro da lugar a la muerte celular por ferroptosis si el medio no es suplementado con un agente reductor como el β-mercaptoetanol. En dichas condiciones, las células deficientes de CD98hc presentan estrés oxidativo y un desequilibrio intracelular de AAs, probablemente debido a la pérdida de CD98hc-LAT1. Consecuentemente, la síntesis proteica y la proliferación son defectuosas en estas células. Además, CD98hc regula el metabolismo energético de la célula, de manera que su eliminación causa una reducción de la captación de glucosa y de las vías metabólicas relacionadas con su metabolismo, como la glucólisis y la ruta de las pentosas fosfato, lo que obliga a las células a utilizar el metabolismo oxidativo mitocondrial para mantener los niveles celulares de ATP. El bloqueo de la ruta de las pentosas fosfato provoca una reducción dramática del contenido de ribosa-5P, lo que se ve reflejado en una disminución global del conjunto de nucleótidos, causando estrés replicativo y alteraciones en la progresión del ciclo celular. Previas consideraciones sugieren que la eliminación de CD98hc es una posible diana terapéutica para el tratamiento anti-tumoral

CD98hc orchestrates amino acid and glucose availability, redox homeostasis and energetic and nucleotide metabolism = CD98hc sostiene la disponibilidad de aminoácidos y glucosa, la homeóstasis óxido-reducción y el metabolismo energético y de nucleótidos

The wide variety of disorders associated with the malfunction of amino acid (AA) transporters reflect the relevant roles they fulfil in human physiology. Heteromeric AA transporters (HATs), one of the eleven families of AA plasma membrane transporters in human cells, are formed by an ancillary heavy subunit, which brings the holotransporter to the plasma membrane and a catalytic light subunit. The heavy subunit CD98hc (also named 4F2hc or SLC3A2) can dimerize with any of six light subunits (LAT1, LAT2, xCT, y+LAT1, y+LAT2 and asc1) that confer substrate specificity to the heterodimer. Cancer cells overexpress CD98hc-transporters in order to meet their increased nutritional and antioxidant demands, since these transporters are crucial to control reactive oxygen species and intracellular AA levels, thus sustaining cell survival and proliferation. CD98hc-xCT is required for cystine uptake, which is rapidly reduced to cysteine inside the cell, a rate-limiting AA in the synthesis of reduced glutathione (GSH). Since GSH is vital for controlling the levels of reactive oxygen species in the cell, loss of CD98hc-xCT in vitro cannot be compensated, leading to cell death by ferroptosis. Supplementation of culture media with β-mercaptoethanol rescues CD98hc-deficient cell survival. Under such conditions cells lacking CD98hc still present increased levels of reactive oxygen species and intracellular AA imbalance, probably due to the loss of CD98hc-LAT1. Consequently, cells lacking CD98hc present alterations in the stress-sensing signalling pathways mTORC1 and Integrated Stress Response- mediated by eIF2α, leading to defective protein synthesis and proliferation. Furthermore, CD98hc regulates the energy metabolism of the cell. Indeed, lack of CD98hc triggers a reduction the glucose uptake and glucose-related metabolic pathways, such as glycolysis and the pentose phosphate pathway, forcing cells to use mitochondrial oxidative metabolism in order to maintain the cellular ATP levels, suggesting a metabolic rewiring. The abrogation of the pentose phosphate pathway flux by CD98hc ablation triggers a dramatic reduction in the levels of ribose-5P, which forms the sugar backbone of all nucleotides. Consequently, these cells present a dramatic reduction in the nucleotide pool, which cause replicative stress, enhanced DNA Damage Response, delayed cell cycle progression during the S-phase and diminished rate of mitosis, all recovered by nucleoside supplementation. In addition, reduction in the intracellular content of the essential branched-chain AAs (leucine, isoleucine and valine) and aromatic AAs (tyrosine, phenylalanine and tryptophan), all transported by CD98hc-LAT1, leads to repressed expression of RRM2, the only enzyme able to catalyse the reduction of ribonucleotides to deoxyribonucleotides, resulting as well in replicative stress. Taking all of these considerations, CD98hc may be a putative target in pathophysiological scenarios, thus, generating drugs against CD98hc could represent a novel option, widening the therapeutic window of cancer therapy by inducing an immediate impairment in: the redox counterbalance capacity, the intracellular AA content for proper protein synthesis and cell proliferation and the pentose phosphate pathway flux required for producing ribose-5P to sustain nucleotide biosynthesis of the cells.La gran variedad de trastornos asociados con el mal funcionamiento de los transportadores de aminoácidos (AA) evidencia la importancia de las funciones que éstos llevan a cabo en la fisiología humana. Los transportadores heteroméricos de AAs (HATs) están formados por una subunidad pesada, que sitúa el heterodímero en la membrana plasmática, y una subunidad ligera que confiere la capacidad catalítica al transportador. La subunidad pesada CD98hc (también denominada 4F2hc o SLC3A2) puede dimerizar las subunidades ligeras LAT1, LAT2, xCT, y + LAT1, y + LAT2 y asc1, confiriendo especificidad de sustrato al heterodímero. Las células tumorales sobreexpresan los transportadores asociados a CD98hc para satisfacer sus demandas nutricionales y antioxidantes, ya que estos transportadores son cruciales para controlar las especies reactivas de oxígeno y los niveles intracelulares de AA, soportando la viabilidad y la proliferación celular. CD98hc-xCT es el responsable de la captación de cistina, necesaria para sintetizar glutatión y por tanto para mantener la homeostasis óxido-reducción. En la célula. Por este motivo, la eliminación de CD98hc-xCT in vitro da lugar a la muerte celular por ferroptosis si el medio no es suplementado con un agente reductor como el β-mercaptoetanol. En dichas condiciones, las células deficientes de CD98hc presentan estrés oxidativo y un desequilibrio intracelular de AAs, probablemente debido a la pérdida de CD98hc-LAT1. Consecuentemente, la síntesis proteica y la proliferación son defectuosas en estas células. Además, CD98hc regula el metabolismo energético de la célula, de manera que su eliminación causa una reducción de la captación de glucosa y de las vías metabólicas relacionadas con su metabolismo, como la glucólisis y la ruta de las pentosas fosfato, lo que obliga a las células a utilizar el metabolismo oxidativo mitocondrial para mantener los niveles celulares de ATP. El bloqueo de la ruta de las pentosas fosfato provoca una reducción dramática del contenido de ribosa-5P, lo que se ve reflejado en una disminución global del conjunto de nucleótidos, causando estrés replicativo y alteraciones en la progresión del ciclo celular. Previas consideraciones sugieren que la eliminación de CD98hc es una posible diana terapéutica para el tratamiento anti-tumoral

Nitroglicerina sublingual: conocimientos al alta hospitalaria

Màster en Infermeria Cardiovascular, Universitat de Barcelona, Escola Universitària d'Infermeria, any: 2011-2012, Director: Joan Maria Estrada Masllorens. Tutora: Sandra Cabrera JaimeLa cardiopatía isquémica es aquella situación en la cual el corazón sufre una lesión, isquemia o necrosis causada por obstrucción de algún o algunos vasos coronarios, produciendo falta de irrigación y oxigenación de los mismos. Ésta enfermedad esta ocasionada por la arterioesclerosis de las arterias coronarias, que son las encargadas de proporcionar la sangre al miocardio. La arterioesclerosis coronaria es un proceso lento de formación de colágeno y acumulación de lípidos y células inflamatorias, causante de la estenosis. El tratamiento más común es antiagregantes, fármacos antianginosos como los nitratos, betabloqueantes, calcioantagonistas y la nitroglicerina sublingual si precis

Conservación de la escultura contemporánea en acero patinable en el exterior: empleo de recubrimientos protectores.

A través de este artículo presentamos los resultados obtenidos en los trabajos preliminares de la tesis doctoral «Sistemas de protección para la escultura contemporánea en acero COR-TEN», que actualmente se encuentra en fase de desarrollo

Expression of the Gene for Autotransporter AutB of Neisseria meningitidis Affects Biofilm Formation and Epithelial Transmigration

Neisseria meningitidis is a Gram-negative bacterium that resides as a commensal in the upper respiratory tract of humans, but occasionally, it invades the host and causes sepsis and/or meningitis. The bacterium can produce eight autotransporters, seven of which have been studied to some detail. The remaining one, AutB, has not been characterized yet. Here, we show that the autB gene is broadly distributed among pathogenic Neisseria spp. The gene is intact in most meningococcal strains. However, its expression is prone to phase variation due to slipped-strand mispairing at AAGC repeats located within the DNA encoding the signal sequence and is switched off in the vast majority of these strains. Moreover, various genetic disruptions prevent autB expression in most of the strains in which the gene is in phase indicating a strong selection against AutB synthesis. We observed that autB is expressed in two of the strains examined and that AutB is secreted and exposed at the cell surface. Functionality assays revealed that AutB synthesis promotes biofilm formation and delays the passage of epithelial cell layers in vitro. We hypothesize that this autotransporter is produced during the colonization process only in specific niches to facilitate microcolony formation, but its synthesis is switched off probably to evade the immune system and facilitate human tissue invasion